RESUMEN
As other European countries, France is experiencing a resurgence of pertussis in 2024. Between 1 January and 31 May 2024, 5,616 (24.9%) positive Bordetella pertussis qPCR tests were identified, following a 3-year period of almost null incidence. Of 67 cultured and whole genome sequenced B. pertussis isolates, 66 produced pertactin and 56 produced FIM2, in contrast to pre-COVID-19 years. One isolate of genotype Bp-AgST4 was resistant to macrolides. Pertussis resurgence may favour isolates that produce FIM2 and pertactin.
Asunto(s)
Antibacterianos , Bordetella pertussis , Macrólidos , Tos Ferina , Bordetella pertussis/genética , Bordetella pertussis/aislamiento & purificación , Bordetella pertussis/efectos de los fármacos , Humanos , Francia/epidemiología , Macrólidos/farmacología , Tos Ferina/epidemiología , Tos Ferina/microbiología , Antibacterianos/farmacología , Farmacorresistencia Bacteriana , Pruebas de Sensibilidad Microbiana , Proteínas de la Membrana Bacteriana Externa/genética , Secuenciación Completa del Genoma , Factores de Virulencia de Bordetella/genética , Genotipo , Adulto , Niño , Incidencia , PreescolarRESUMEN
Between March and October 2022, a peak of detection of Bordetella parapertussis by qPCR, real-time PCR was observed in France.Hypothesis/Gap Statement. Whether this peak was due to resurgence from previous circulating lineages or reintroduction into the country was unknown.Objective. The objective of this study is to understand B. parapertussis-transient increase observed in France in 2022 whereas it had virtually stopped being reported since the start of the COVID-19 pandemic in 2020.Methods. We analysed real-time PCR (qPCR) data from the two largest French outpatient laboratories performing whooping cough diagnosis and characterized all B. parapertussis isolates collected in the 2016-2022 period by the French National Reference Centre for Whooping Cough.Results. Microbiological analyses reveal that 13 of 18 bacterial isolates collected in 2022 produce the vaccine antigen pertactin, whereas none of the 22 isolates collected in the 2016-2021 period did.Conclusion. We hypothesize a re-introduction of B. parapertussis from regions of the world where whole-cell vaccines are still in use.
Asunto(s)
Bordetella parapertussis , Tos Ferina , Francia/epidemiología , Humanos , Bordetella parapertussis/genética , Bordetella parapertussis/aislamiento & purificación , Tos Ferina/epidemiología , Tos Ferina/microbiología , Reacción en Cadena en Tiempo Real de la Polimerasa , Proteínas de la Membrana Bacteriana Externa/genética , Infecciones por Bordetella/microbiología , Infecciones por Bordetella/epidemiología , Niño , Preescolar , Adulto , Factores de Virulencia de Bordetella/genética , Femenino , COVID-19/epidemiología , Adolescente , Lactante , Masculino , Adulto JovenRESUMEN
Bordetella pertussis persists inside host cells, and virulence factors are crucial for intracellular adaptation. The regulation of B. pertussis virulence factor transcription primarily occurs through the modulation of the two-component system (TCS) known as BvgAS. However, additional regulatory systems have emerged as potential contributors to virulence regulation. Here, we investigate the impact of BP1092, a putative TCS histidine kinase that shows increased levels after bacterial internalization by macrophages, on B. pertussis proteome adaptation under nonmodulating (Bvg+) and modulating (Bvg-) conditions. Using mass spectrometry, we compare B. pertussis wild-type (wt), a BP1092-deficient mutant (ΔBP1092), and a ΔBP1092 trans-complemented strain under both conditions. We find an altered abundance of 10 proteins, including five virulence factors. Specifically, under nonmodulating conditions, the mutant strain showed decreased levels of FhaB, FhaS, and Cya compared to the wt. Conversely, under modulating conditions, the mutant strain exhibited reduced levels of BvgA and BvgS compared to those of the wt. Functional assays further revealed that the deletion of BP1092 gene impaired B. pertussis ability to survive within human macrophage THP-1 cells. Taken together, our findings allow us to propose BP1092 as a novel player involved in the intricate regulation of B. pertussis virulence factors and thus in adaptation to the intracellular environment. The data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the data set identifier PXD041940.
Asunto(s)
Proteínas Bacterianas , Bordetella pertussis , Histidina Quinasa , Bordetella pertussis/patogenicidad , Bordetella pertussis/genética , Histidina Quinasa/metabolismo , Histidina Quinasa/genética , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Virulencia/genética , Regulación Bacteriana de la Expresión Génica , Macrófagos/microbiología , Humanos , Proteoma , Factores de Virulencia de Bordetella/genética , Factores de Virulencia de Bordetella/metabolismo , Factores de Virulencia/genética , Factores de Virulencia/metabolismo , Viabilidad MicrobianaRESUMEN
Bordetella species that cause respiratory infections in mammals include B. pertussis, which causes human whooping cough, and B. bronchiseptica, which infects nearly all mammals. Both bacterial species produce filamentous hemagglutinin (FhaB) and adenylate cyclase toxin (ACT), prominent surface-associated and secreted virulence factors that contribute to persistence in the lower respiratory tract by inhibiting clearance by phagocytic cells. FhaB and ACT proteins interact with themselves, each other, and host cells. Using immunoblot analyses, we showed that ACT binds to FhaB on the bacterial surface before it can be detected in culture supernatants. We determined that SphB1, a surface protease identified based on its requirement for FhaB cleavage, is also required for ACT cleavage, and we determined that the presence of ACT blocks SphB1-dependent and -independent cleavage of FhaB, but the presence of FhaB does not affect SphB1-dependent cleavage of ACT. The primary SphB1-dependent cleavage site on ACT is proximal to ACT's active site, in a region that is critical for ACT activity. We also determined that FhaB-bound ACT on the bacterial surface can intoxicate host cells producing CR3, the receptor for ACT. In addition to increasing our understanding of FhaB, ACT, and FhaB-ACT interactions on the Bordetella surface, our data are consistent with a model in which FhaB functions as a novel toxin delivery system by binding to ACT and allowing its release upon binding of ACT to its receptor, CR3, on phagocytic cells.IMPORTANCEBacteria need to control the variety, abundance, and conformation of proteins on their surface to survive. Members of the Gram-negative bacterial genus Bordetella include B. pertussis, which causes whooping cough in humans, and B. bronchiseptica, which causes respiratory infections in a broad range of mammals. These species produce two prominent virulence factors, the two-partner secretion (TPS) effector FhaB and adenylate cyclase toxin (ACT), that interact with themselves, each other, and host cells. Here, we determined that ACT binds FhaB on the bacterial surface before being detected in culture supernatants and that ACT bound to FhaB can be delivered to eukaryotic cells. Our data are consistent with a model in which FhaB delivers ACT specifically to phagocytic cells. This is the first report of a TPS system facilitating the delivery of a separate polypeptide toxin to target cells and expands our understanding of how TPS systems contribute to bacterial pathogenesis.
Asunto(s)
Toxina de Adenilato Ciclasa , Fagocitos , Factores de Virulencia de Bordetella , Toxina de Adenilato Ciclasa/metabolismo , Toxina de Adenilato Ciclasa/genética , Fagocitos/metabolismo , Fagocitos/microbiología , Factores de Virulencia de Bordetella/metabolismo , Factores de Virulencia de Bordetella/genética , Humanos , Bordetella pertussis/metabolismo , Bordetella pertussis/genética , Adhesinas Bacterianas/metabolismo , Adhesinas Bacterianas/genética , Bordetella bronchiseptica/metabolismo , Bordetella bronchiseptica/genética , Unión Proteica , AnimalesRESUMEN
The Gram-negative pathogenic bacterium Bordetella bronchiseptica is a respiratory pathogen closely related to Bordetella pertussis, the causative agent of whooping cough. Despite sharing homologous virulence factors, B. bronchiseptica infects a broad range of mammalian hosts, including some experimental animals, whereas B. pertussis is strictly adapted to humans. Therefore, B. bronchiseptica is often used as a representative model to explore the pathogenicity of Bordetella in infection experiments with laboratory animals. Although Bordetella virulence factors, including toxins and adhesins have been studied well, our recent study implied that unknown virulence factors are involved in tracheal colonization and infection. Here, we investigated bacterial genes contributing to tracheal colonization by high-throughput transposon sequencing (Tn-seq). After the screening, we picked up 151 candidate genes of various functions and found that a rpoN-deficient mutant strain was defective in tracheal colonization when co-inoculated with the wild-type strain. rpoN encodes σ54 , a sigma factor that regulates the transcription of various genes, implying its contribution to various bacterial activities. In fact, we found RpoN of B. bronchiseptica is involved in bacterial motility and initial biofilm formation. From these results, we propose that RpoN supports bacterial colonization by regulating various bacteriological functions.
Asunto(s)
Infecciones por Bordetella , Bordetella bronchiseptica , Bordetella , Animales , Humanos , Bordetella bronchiseptica/genética , ARN Polimerasa Sigma 54 , Bordetella pertussis/genética , Factores de Virulencia de Bordetella/genética , Factores de Virulencia/genética , MamíferosRESUMEN
There has been a resurgence in the number of pertussis cases in Brazil and around the world. Here, the genome of a clinical Bordetella pertussis strain (Bz181) that was recently isolated in Brazil is reported. Analysis of the virulence-associated genes defining the pre- and post-vaccination lineages revealed the presence of the prn2-ptxS1A-fim3B-ptxP3 allelic profile in Bz181, which is characteristic of the current pandemic lineage. A putative metallo-β-lactamase gene presenting all of the conserved zinc-binding motifs that characterise the catalytic site was identified, in addition to a multidrug efflux pump of the RND family that could confer resistance to erythromycin, which is the antibiotic of choice for treating pertussis disease.